朱桔Mn-SOD的纯化鉴定及浓度梯度胶电泳在其中的应用-植物研究.PDF

24 　2 　　　　　　　　　　　　　　　　　　　　　　 2004 　4 Vol.24　No.2　　　　　　　　　　　　BULLETIN OF BOTANICAL RESEARCH April 　2004 Mn-SOD 、 程光宇　吴国荣　储慧君 (, 　210097) 　　朱桔叶中存在Mn-SOD、Fe-SOD 和Cu n-SOD 三种类型, 在 10%的PAGE 中有三条活性 带, 其中Mn-SOD 的R 值大于Fe-SOD 与Cu n-SOD 的R 值相同;在4%～35%的梯度胶电泳中 f 1 f 有四条活性带, 而Mn-SOD 的R 值 小, 表明该酶带有较高的电荷密度。Mn-SOD 占总活性的f 20%左右, 已被纯化到均 一程度。该酶的比活性为 1 249 U/mg, 分子量和亚基分子量分别为 54.0 kD和26.6 kD, 在紫外区 大吸收值为280 nm, 在 95 ℃处理 15 min 仍保留了46%的酶活 性, 等电点为5.06。该酶的活性不被KCN 、H O 抑制, 但对1%SDS 和氯仿-乙醇液敏感。 2 2 　朱桔;Mn-SOD;纯化;性质;电荷密度 Purification and di fferentiation of Mn-SOD from Citrus ery throsa and application of gel concentration gradient electrophoresis CHENG Guang-Yu　WU Guo-Rong 　CHU Hui-Jun (School of Life Science, Nanjing Normal University, Nanjing 　210097) Abstract 　There were Mn-SOD, Fe-SOD and Cu n-SOD in the crude extract of Citr us ery t hrosa Tanaka leaves.In a 10%PAGE three active bands were seen, and R value of Mn-SOD was big thanf that of Fe-SOD and was same as that of Cu n-SOD .While in a 4%～35%gel concentration gradi- 1 ent electrophoresis, four active bands were seen and Mn-SOD s R value was the smallest .Thisf means that the molecular weight of Mn-SOD was the biggest and it had a high charge density .Mn- SOD, which occupied about 20%of the total SOD activity, was purified to homogeneity by means of ammonium sulfate precipitation, Sephadex G-100 gel filtration, DEAE-Sepharose and CM52 chro- matography etc.Its specific activity was 1 249 U/mg.Its molecular an